Establishment of Enteritis Model in Mice Infected with Enterotoxigenic Escherichia coli and Its Molecular Mechanism

WU Yue; WANG Yimeng; WANG Mengmeng; ZHOU Jiangtao; LIU Xuejiao; LI Haihua; QIAO Jiayun

doi:10.3969/j.issn.1006-267x.2021.10.041

Chinese Journal of Animal Nutrition >

2021 , Vol. 33 >Issue 10: 5817 - 5826

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.10.041

MOLECULAR AND CELLULAR NUTRITION

Establishment of Enteritis Model in Mice Infected with Enterotoxigenic Escherichia coli and Its Molecular Mechanism

WU Yue ,
WANG Yimeng ,
WANG Mengmeng ,
ZHOU Jiangtao ,
LIU Xuejiao ,
LI Haihua ,
QIAO Jiayun

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1. Tianjin Key Laboratory of Agriculture Animal Breeding and Healthy Husbandry, College of Animal Science and Animal Medicine, Tianjin Agricultural University, Tianjin 300384, China;
2. Tianjin Key Laboratory of Conservation and Utilization of Animal Diversity, College of Life Sciences, Tianjin Normal University, Tianjin 300387, China

Received date: 2021-03-07

Online published: 2021-10-16

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Abstract

The aim of this study was to establish an enterotoxigenic Escherichia coli (ETEC) infection model of mouse enteritis, and to reveal its molecular mechanism. A total of 120 mice were randomly divided into 4 groups with 30 mice in each group, and there were 5 replicates per group and 6 replicates per replicate. The mice in low, medium and high dose experimental groups were given 1×10⁵, 1×10⁶ and 1×10⁷ CFU/mice by gastric gavage, respectively, while those in control group was given equal volume of normal saline by gavage, with the injection dose of 0.2 mL in each group. Clinical manifestations of mice were observed and weighed at 24, 48, 72, 96 and 120 h after challenge, respectively. Meanwhile, 6 mice were randomly selected from each treatment group and their blood and tissue samples were collected. The contents of C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukin-8 (IL-8) in serum were detected by enzyme-linked immunosorbent assay (ELISA). The pathological changes of jejunum, liver and spleen of mice were observed by tissue sections and hematoxylin-eosin staining (HE staining). The mRNA relative expression levels of Toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB p65), myeloid differentiation factor 88 (MyD88) and recombinant B-cell (BCl3) were detected by fluorescence quantitative PCR. The results showed as follows:compared with the control group, challenged mice were depressed and eating less, the body weight of the three experimental groups was significantly decreased at 72 and 96 h after challenge (P<0.01 or P<0.05), intestinal, liver and spleen tissue sections showed obvious inflammatory reaction; the contents of CRP, TNF-α and IL-8 in serum of test groups were significantly increased at 48 and 72 h after challenge (P<0.01). The mRNA relative expression levels of TLR4, NF-κB p65 and MyD88 in jejunum of experimental groups were significantly increased at 48, 72 and 96 h after challenge (P<0.01). The mRNA relative expression levels of BCl3 in jejunum of test groups was significantly increased at 24 and 120 h after challenge (P<0.01 or P<0.05), and was significantly decreased at 72 h after challenge (P<0.01). In conclusion, this study successfully establishes the enterotoxigenic Escherichia coli infected mouse enteritis model, and the pathogenic bacteria may induce the production of inflammatory related factors and inflammatory cell infiltration in tissues through TLR4/NF-κB signaling pathway, thus causing inflammatory response in mice.

Key words： Escherichia coli; TLR4/NF-κ; B signaling pathway; inflammatory response; colitis in mice

Cite this article

WU Yue , WANG Yimeng , WANG Mengmeng , ZHOU Jiangtao , LIU Xuejiao , LI Haihua , QIAO Jiayun . Establishment of Enteritis Model in Mice Infected with Enterotoxigenic Escherichia coli and Its Molecular Mechanism[J]. Chinese Journal of Animal Nutrition, 2021 , 33(10) : 5817 -5826 . DOI: 10.3969/j.issn.1006-267x.2021.10.041

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